Visualization of lysosomal membrane proteins by cryo electron tomography.

IF 15.7 1区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Nature Communications Pub Date : 2025-10-17 DOI:10.1038/s41467-025-64314-0

Bridget M McVeigh,José J De Jesús-Pérez,Dirk H Siepe,Prerana Gogoi,Shrawan Kumar Mageswaran,Marian Kalocsay,Elaine M Mihelc,Vera Y Moiseenkova-Bell

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Abstract

Lysosomes are essential organelles for cellular homeostasis and signaling, with dysfunction linked to neurological disorders, lysosomal storage diseases, and cancer. While proteomics has advanced our understanding of lysosomal composition, the structural characterization of lysosomal membrane proteins in their native environment remains a significant challenge. Here, we developed a cryo electron tomography workflow to visualize lysosomal membrane proteins within intact, native lysosomal membranes. We isolated endolysosomes by independently targeting two lysosomal membrane proteins, transient receptor potential mucolipin 1 and transmembrane protein 192, enriching organelles that exhibited the expected morphology and proteomic composition of the endolysosomal system. Sub-tomogram averaging enabled the structural refinement of key membrane and membrane-associated proteins, including V-ATPase, Flotillin, and Clathrin, directly within the lysosomal membrane, revealing their heterogeneous distribution across endolysosomal organelles. By integrating proteomics with structural biology, our workflow establishes a powerful platform for studying lysosomal membrane protein function in health and disease, paving the way for future discoveries in membrane-associated lysosomal mechanisms.

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溶酶体膜蛋白的低温电子断层成像。

溶酶体是细胞内稳态和信号传导的重要细胞器，其功能障碍与神经系统疾病、溶酶体贮积病和癌症有关。虽然蛋白质组学提高了我们对溶酶体组成的理解，但溶酶体膜蛋白在其天然环境中的结构表征仍然是一个重大挑战。在这里，我们开发了一种低温电子断层扫描工作流程来可视化完整的天然溶酶体膜内的溶酶体膜蛋白。我们通过独立靶向两种溶酶体膜蛋白，瞬时受体电位粘磷脂1和跨膜蛋白192分离出内溶酶体，富集了表现出内溶酶体系统预期形态和蛋白质组学组成的细胞器。亚断层扫描平均能够直接在溶酶体膜内对关键的膜和膜相关蛋白进行结构细化，包括v - atp酶、Flotillin和Clathrin，揭示了它们在溶酶体细胞器中的异质性分布。通过将蛋白质组学与结构生物学相结合，我们的工作流程为研究溶酶体膜蛋白在健康和疾病中的功能建立了一个强大的平台，为未来发现膜相关溶酶体机制铺平了道路。

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来源期刊

Nature Communications Biological Science Disciplines-

CiteScore

24.90

自引率

2.40%

发文量

6928

审稿时长

3.7 months

期刊介绍： Nature Communications, an open-access journal, publishes high-quality research spanning all areas of the natural sciences. Papers featured in the journal showcase significant advances relevant to specialists in each respective field. With a 2-year impact factor of 16.6 (2022) and a median time of 8 days from submission to the first editorial decision, Nature Communications is committed to rapid dissemination of research findings. As a multidisciplinary journal, it welcomes contributions from biological, health, physical, chemical, Earth, social, mathematical, applied, and engineering sciences, aiming to highlight important breakthroughs within each domain.